ANTIGEN RETRIEVAL PROTOCOL FOR PARAFFIN-EMBEDDED TISSUE

In the previous section on DEPARAFFINIZATION PRIOR TO IMMUNOHISTOCHEMISTRY STAINING, we looked at deparaffinization and rehydration of paraffin-embedded tissue. In this section, we look at epitope unmasking protocol. Deparaffinization is an important process in the immunohistochemistry staining process; and it must be done prior to immunohistochemistry, especially for paraffin-embedded tissue samples. Deparaffinization is simply defined […]

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Immunology & Immune System, , , , ,

ANTIBODY SELECTION AND STAINING FOR IMMUNOHISTOCHEMISTRY

We have looked at deparaffinization and rehydration of paraffin-embedded tissue, as well as antigen-retrieval protocol and permeabilization and blocking non-specific binding on the sectioned tissue sample on the slide. In this section, we look briefly on antibody selection and antibody staining for immunohistochemistry. The selection of antibody for immunohistochemistry analysis is important for the overall

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Immunology & Immune System, , ,

ADENOVIRUS INFECTION

Adenoviruses are found in the viral family known as Adenoviridae. They are non-enveloped viruses with an icosahedral nucleocapsid. Adenoviruses are DNA viruses; and they possess a double stranded DNA molecule. Adenoviruses are a group of viruses that typically cause respiratory illnesses, such as a common cold, conjunctivitis, croup, bronchitis, or pneumonia. In children, adenoviruses usually

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Virology, , ,

NITROCEFIN TEST FOR BETA-LACTAMASE PRODUCTION

The production of beta-lactamase enzymes by Gram negative bacteria including E. coli, P. aeruginosa and Klebsiella species is an important mechanism by which the organisms evade both in vivo and in vitro antimicrobial onslaught. Beta-lactamase production by resistant test isolates can be evaluated for beta-lactamase production using the Nitrocefin test sticks (Oxoid, UK). The nitrocefin

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Antibiotic Resistance / Antimicrobial Resistance (AMR), Pharmaceutical Microbiology, , ,

WAYS TO SEPARATE DNA FROM OTHER CELLULAR MATERIALS: BINDING AND WASHING

When phenol or a mixture of phenol:chloroform is mixed with a  cell Lysate, two phases form: an aqueous and an organic phase. The polar DNA partitions into the polar or “aqueous phase”, and denatured proteins and other cellular debris partition into the organic phase. Salts are thought to dehydrate proteins, causing them to become less

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Biotechnology, Molecular Microbiology, ,

Vancomycin resistance in Enterococci

Enterococcus (plural: Enterococci) is a group of bacteria that is normally found in the intestines of humans and many animals. Two species of Enterococci, which are found in the body of healthy individuals, are Enterococcus faecalis and Enterococcus faecium. Both E. faecalis and E. faecium are associated with human diseases; and they are mostly involved

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Antibiotic Resistance / Antimicrobial Resistance (AMR), Pharmaceutical Microbiology, , , , , ,

TYPES OF SOLID SUPPORT FOR BINDING DNA

The types of solid support for binding DNA are as follows: 1. Silica Matrix Silica Matrix is a solid support for binding DNA. DNA binds to silica in the presence of high concentrations of chaotropic salts (e.g., guanidium HCL), but protein does not. Salts can be removed with an alcohol-based wash and DNA eluted in

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Molecular Microbiology, , , ,

What are ESBLs?

ESBLs (extended spectrum beta-lactamases) are enzymes that mediate resistance to extended-spectrum (third generation) cephalosporins (e.g., ceftazidime, cefotaxime, and ceftriaxone) and monobactams (e.g., aztreonam) but do not affect cephamycins (e.g., cefoxitin and cefotetan) or carbapenems (e.g., meropenem or imipenem). Why should hospital laboratories and hospitals be concerned about detecting ESBL-producing bacteria? The presence of an ESBL-producing

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Pharmaceutical Microbiology, Antibiotic Resistance / Antimicrobial Resistance (AMR), , ,

What is a stem cell?

A stem cell is a cell with the unique ability to develop into specialised cell types in the body. In the future they may be used to replace cells and tissues that have been damaged or lost due to disease. Figure 1 is an illustration showing a stem cell giving rise to more stem cells or specialised

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Cell Culture Notes, Immunology & Immune System, , , , , ,